P-selectin and E-selectin. Distinct but overlapping leukocyte ligand specificities.
P-selectin and E-selectin. Distinct but overlapping leukocyte ligand specificities.
复制标题
DOI：
10.1016/s0021-9258(19)49881-5
复制
发表时间：
1992-06
期刊：
影响因子：
--
通讯作者：
Glenn R. Larsen;Dianne S. Sako;T. J. Ahern;M. Shaffer;John K. Erban;S. Sajer;R. Gibson;Denisa D. Wagner;Bruce Furie;Bruce Furie
中科院分区：
文献类型：
--
作者：
Glenn R. Larsen;Dianne S. Sako;T. J. Ahern;M. Shaffer;John K. Erban;S. Sajer;R. Gibson;Denisa D. Wagner;Bruce Furie;Bruce Furie
血小板和内皮细胞上的 P-选择素和内皮细胞上的 E-选择素是白细胞受体，可识别中性粒细胞和单核细胞上的谱系特异性碳水化合物。这些受体的拟议配体包含 Le(x) 核心和唾液酸。由于其他研究人员表明 E-选择素和 P-选择素均与唾液酸化 Le(x) 结合，因此我们评估了 E-选择素和 P-选择素是否识别白细胞上相同的反受体。研究了 HL60 细胞与表达 P-选择素或 E-选择素的中国仓鼠卵巢 (CHO) 细胞的相互作用。为了确定除了唾液酸 Le(x) 之外是否还需要蛋白质成分来识别 P-选择素或 E-选择素，用蛋白酶（包括胰凝乳蛋白酶、弹性蛋白酶、蛋白酶 Glu-C、无花果蛋白酶、木瓜蛋白酶或嗜热菌蛋白酶）消化 HL60 细胞或中性粒细胞。用这些蛋白酶处理的细胞结合 E-选择素，但不结合 P-选择素。 HL60细胞的岩藻糖苷酶或神经氨酸酶处理显着降低了与表达E-选择素和P-选择素的CHO细胞的结合。 HL60 细胞在衣霉素中的生长抑制了这些细胞支持 P-选择素介导的结合的能力，并在较小程度上抑制了 E-选择素介导的结合。纯化的P-选择素抑制CHO:P-选择素与HL60细胞的结合，但不完全抑制CHO:E-选择素与HL60细胞的结合。然而，纯化的可溶性E-选择素同等且完全地抑制CHO:P-选择素和CHO:E-选择素与HL60细胞的结合。不能结合E-选择素或P-选择素的COS细胞在用α-1,3-岩藻糖基转移酶或α-1,3/1,4-岩藻糖基转移酶转染后结合E-选择素但不结合P-选择素。类似地，表达唾液酸Le(x)的LEC 11细胞结合E-选择素但不结合表达P-选择素的CHO细胞。黑接骨木凝集素对 sialyl-2,6 beta Gal/GalNAc 连接具有特异性，可抑制 P-选择素但不抑制 E-选择素与 HL60 细胞的结合。尽管唾液酸和Le(x)是P-选择素配体和E-选择素配体的组分，但这些结果表明配体是相关的，具有重叠的特异性，但结构上不同。在 P-选择素配体上的唾液酸-2,6 beta Gal 结构附近含有唾液酸 Le(x) 的蛋白质成分可能有助于其对 P-选择素的特异性。
P-selectin on platelets and endothelial cells and E-selectin on endothelial cells are leukocyte receptors that recognize lineage-specific carbohydrates on neutrophils and monocytes. The proposed ligands for these receptors contain the Le(x) core and sialic acid. Since other investigators have shown that both E-selectin and P-selectin bind to sialylated Le(x), we evaluated whether E-selectin and P-selectin recognize the same counter-receptor on leukocytes. The interaction of HL60 cells with Chinese hamster ovary (CHO) cells expressing P-selectin or E-selectin was studied. To determine whether a protein component is required in addition to sialyl Le(x) for either P-selectin or E-selectin recognition, HL60 cells or neutrophils were digested with proteases, including chymotrypsin, elastase, proteinase Glu-C, ficin, papain, or thermolysin. Cells treated with these proteases bound E-selectin but not P-selectin. Fucosidase or neuraminidase treatment of HL60 cells markedly decreased binding to both E-selectin- and P-selectin-expressing CHO cells. Growth of HL60 cells in tunicamycin inhibited the ability of these cells to support P-selectin-mediated binding and, to a lesser extent, E-selectin-mediated binding. Purified P-selectin inhibited CHO:P-selectin binding to HL60 cells, but incompletely inhibited CHO:E-selectin binding to HL60 cells. However, purified soluble E-selectin inhibited CHO:P-selectin and CHO:E-selectin binding to HL60 cells equivalently and completely. COS cells, unable to bind to E-selectin or P-selectin, bound E-selectin but not P-selectin upon transfection with alpha-1,3-fucosyltransferase or alpha-1,3/1,4-fucosyltransferase. Similarly, LEC 11 cells expressing sialyl Le(x) bound E-selectin- but not P-selectin-expressing CHO cells. Sambucus nigra lectin, specific for the sialyl-2,6 beta Gal/GalNAc linkage, inhibited P-selectin but not E-selectin binding to HL60 cells. Although sialic acid and Le(x) are components of the P-selectin ligand and the E-selectin ligand, these results indicate that the ligands are related, having overlapping specificities, but are structurally distinct. A protein component containing sialyl Le(x) in proximity to sialyl-2,6 beta Gal structures on the P-selectin ligand may contribute to its specificity for P-selectin.